{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,24]],"date-time":"2026-03-24T18:54:25Z","timestamp":1774378465160,"version":"3.50.1"},"reference-count":58,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2019,9,5]],"date-time":"2019-09-05T00:00:00Z","timestamp":1567641600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Fabry disease (FD) is caused by mutations in the GLA gene that encodes lysosomal \u03b1-galactosidase-A (\u03b1-gal-A). A number of pathogenic mechanisms have been proposed and these include loss of mitochondrial respiratory chain activity. For FD, gene therapy is beginning to be applied as a treatment. In view of the loss of mitochondrial function reported in FD, we have considered here the impact of loss of mitochondrial respiratory chain activity on the ability of a GLA lentiviral vector to increase cellular \u03b1-gal-A activity and participate in cross correction. Jurkat cells were used in this study and were exposed to increasing viral copies. Intracellular and extracellular enzyme activities were then determined; this in the presence or absence of the mitochondrial complex I inhibitor, rotenone. The ability of cells to take up released enzyme was also evaluated. Increasing transgene copies was associated with increasing intracellular \u03b1-gal-A activity but this was associated with an increase in Km. Release of enzyme and cellular uptake was also demonstrated. However, in the presence of rotenone, enzyme release was inhibited by 37%. Excessive enzyme generation may result in a protein with inferior kinetic properties and a background of compromised mitochondrial function may impair the cross correction process.<\/jats:p>","DOI":"10.3390\/ijms20184349","type":"journal-article","created":{"date-parts":[[2019,9,6]],"date-time":"2019-09-06T02:59:22Z","timestamp":1567738762000},"page":"4349","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Inhibition of Mitochondrial Complex I Impairs Release of \u03b1-Galactosidase by Jurkat Cells"],"prefix":"10.3390","volume":"20","author":[{"given":"Jonathan","family":"Lambert","sequence":"first","affiliation":[{"name":"Enzyme Unit Great Ormond Street Hospital, London WC1N 3JH, UK"},{"name":"University College London Great Ormond Street Institute of Child Health London, London WC1N 1EH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steven","family":"Howe","sequence":"additional","affiliation":[{"name":"University College London Great Ormond Street Institute of Child Health London, London WC1N 1EH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahad","family":"Rahim","sequence":"additional","affiliation":[{"name":"University College London School of Pharmacy, University College London, London WC1N 1AX, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Derek","family":"Burke","sequence":"additional","affiliation":[{"name":"Enzyme Unit Great Ormond Street Hospital, London WC1N 3JH, UK"},{"name":"University College London Great Ormond Street Institute of Child Health London, London WC1N 1EH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9906-0200","authenticated-orcid":false,"given":"Simon","family":"Heales","sequence":"additional","affiliation":[{"name":"Enzyme Unit Great Ormond Street Hospital, London WC1N 3JH, UK"},{"name":"University College London Great Ormond Street Institute of Child Health London, London WC1N 1EH, UK"},{"name":"Neurometabolic Unit, National Hospital, London WC1N 3BG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2019,9,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3148","DOI":"10.1016\/S0021-9258(18)51888-3","article-title":"Fabrys Disease\u2014Classification as a Sphingolipidosis and Partial Characterization of a Novel Glycolipid","volume":"238","author":"Sweeley","year":"1963","journal-title":"J. Biol. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1056\/NEJM196705252762101","article-title":"Enzymatic defect in Fabry\u2019s disease. Ceramidetrihexosidase deficiency","volume":"276","author":"Brady","year":"1967","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/j.jmb.2004.01.035","article-title":"The molecular defect leading to Fabry disease: Structure of human alpha-galactosidase","volume":"337","author":"Garman","year":"2004","journal-title":"J. Mol. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1002\/(SICI)1521-3773(19990601)38:11<1532::AID-ANIE1532>3.0.CO;2-U","article-title":"Sphingolipids\u2014Their metabolic pathways and the pathobiochemistry of neurodegenerative diseases","volume":"38","author":"Kolter","year":"1999","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1159\/000132028","article-title":"Regional localization of alpha-galactosidase (GLA) to Xpter\u2192q22, hexosaminidase B (HEXB) to 5q13\u2192qter, and arylsulfatase B (ARSB) to 5pter\u2192q13","volume":"38","author":"Fox","year":"1984","journal-title":"Cytogenet. Cell Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4859","DOI":"10.1073\/pnas.83.13.4859","article-title":"Human alpha-galactosidase A: Nucleotide sequence of a cDNA clone encoding the mature enzyme","volume":"83","author":"Bishop","year":"1986","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2743","DOI":"10.1001\/jama.285.21.2743","article-title":"Enzyme replacement therapy in Fabry disease: A randomized controlled trial","volume":"285","author":"Schiffmann","year":"2001","journal-title":"JAMA"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1056\/NEJM200107053450102","article-title":"Safety and efficacy of recombinant human alpha-galactosidase A-replacement therapy in Fabry\u2019s disease","volume":"345","author":"Eng","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1007\/s40265-016-0607-y","article-title":"Migalastat: First Global Approval","volume":"76","author":"Markham","year":"2016","journal-title":"Drugs"},{"key":"ref_10","first-page":"CD006663","article-title":"Enzyme replacement therapy for Anderson-Fabry disease","volume":"7","author":"Gomaa","year":"2016","journal-title":"Cochrane Database Syst. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1136\/jmedgenet-2015-103486","article-title":"Time to treatment benefit for adult patients with Fabry disease receiving agalsidase beta: Data from the Fabry Registry","volume":"53","author":"Ortiz","year":"2016","journal-title":"J. Med. Genet."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1186\/1750-1172-8-47","article-title":"Long term enzyme replacement therapy for Fabry disease: Effectiveness on kidney, heart and brain","volume":"8","author":"Rombach","year":"2013","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"S58","DOI":"10.1016\/j.ymgme.2015.12.295","article-title":"Phenotype of Fabry disease in patients with mutations amenable to migalastat","volume":"117","author":"Hughes","year":"2016","journal-title":"Mol. Genet. Metab."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1002\/cpt.790","article-title":"Lucerastat, an Iminosugar for Substrate Reduction Therapy: Tolerability, Pharmacodynamics, and Pharmacokinetics in Patients With Fabry Disease on Enzyme Replacement","volume":"103","author":"Oder","year":"2018","journal-title":"Clin. Pharmacol. Ther."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"949","DOI":"10.1126\/science.175.4025.949","article-title":"Gene therapy for human genetic disease?","volume":"175","author":"Friedmann","year":"1972","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.omtm.2017.05.003","article-title":"Lentivector Iterations and Pre-Clinical Scale-Up\/Toxicity Testing: Targeting Mobilized CD34(+) Cells for Correction of Fabry Disease","volume":"5","author":"Huang","year":"2017","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2676","DOI":"10.1073\/pnas.051634498","article-title":"Adeno-associated viral vector-mediated gene transfer results in long-term enzymatic and functional correction in multiple organs of Fabry mice","volume":"98","author":"Jung","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7917","DOI":"10.1073\/pnas.93.15.7917","article-title":"Correction in trans for Fabry disease: Expression, secretion and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector","volume":"93","author":"Medin","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"16909","DOI":"10.1073\/pnas.0407572101","article-title":"Bioluminescent imaging of a marking transgene and correction of Fabry mice by neonatal injection of recombinant lentiviral vectors","volume":"101","author":"Yoshimitsu","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1038\/sj.gt.3302839","article-title":"Efficient correction of Fabry mice and patient cells mediated by lentiviral transduction of hematopoietic stem\/progenitor cells","volume":"14","author":"Yoshimitsu","year":"2007","journal-title":"Gene Ther."},{"key":"ref_21","first-page":"1931","article-title":"Circulating alpha-galactosidase A derived from transduced bone marrow cells: Relevance for corrective gene transfer for Fabry disease","volume":"10","author":"Takenaka","year":"1999","journal-title":"Hum. Gene"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1038\/nrm3565","article-title":"Signals from the lysosome: A control centre for cellular clearance and energy metabolism","volume":"14","author":"Settembre","year":"2013","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1093\/eurjhf\/hfq211","article-title":"Cardiac energy metabolism is disturbed in Fabry disease and improves with enzyme replacement therapy using recombinant human galactosidase A","volume":"13","author":"Machann","year":"2011","journal-title":"Eur. J. Heart Fail."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.ijcard.2009.03.045","article-title":"Association between cardiac energy metabolism and gain of left ventricular mass in Fabry disease","volume":"144","author":"Palecek","year":"2010","journal-title":"Int. J. Cardiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Liebau, M.C., Braun, F., H\u00f6pker, K., Weitbrecht, C., Bartels, V., M\u00fcller, R.-U., Brodesser, S., Saleem, M.A., Benzing, T., and Schermer, B. (2013). Dysregulated autophagy contributes to podocyte damage in Fabry\u2019s disease. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0063506"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/2051-5960-2-20","article-title":"Autophagy-lysosome pathway associated neuropathology and axonal degeneration in the brains of alpha-galactosidase A-deficient mice","volume":"2","author":"Nelson","year":"2014","journal-title":"Acta Neuropathol. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.ymgme.2008.06.016","article-title":"Globotriaosylceramide induces oxidative stress and up-regulates cell adhesion molecule expression in Fabry disease endothelial cells","volume":"95","author":"Shen","year":"2008","journal-title":"Mol. Genet. Metab."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1760","DOI":"10.1016\/j.humpath.2015.07.017","article-title":"Increased oxidative stress contributes to cardiomyocyte dysfunction and death in patients with Fabry disease cardiomyopathy","volume":"46","author":"Chimenti","year":"2015","journal-title":"Hum. Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1111\/j.1749-6632.2002.tb02094.x","article-title":"Role of mitochondria in oxidative stress and aging","volume":"959","author":"Lenaz","year":"2002","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/S0065-2423(09)49003-6","article-title":"Biochemical basis of Fabry disease with emphasis on mitochondrial function and protein trafficking","volume":"49","author":"Das","year":"2009","journal-title":"Adv. Clin. Chem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.ymgme.2004.01.011","article-title":"Fabry disease: Reduced activities of respiratory chain enzymes with decreased levels of energy-rich phosphates in fibroblasts","volume":"82","author":"Lucke","year":"2004","journal-title":"Mol. Genet. Metab."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1016\/j.beem.2012.05.003","article-title":"Cellular and molecular mechanisms of mitochondrial function","volume":"26","author":"Osellame","year":"2012","journal-title":"Best Pract. Res. Clin. Endocrinol. Metab."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1007\/BF00918060","article-title":"Biochemical characteristics of ATP-induced secretion of lysosomal enzymes from rabbit polymorphonuclear leukocytes","volume":"1","author":"Becker","year":"1975","journal-title":"Inflamm"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.redox.2016.12.001","article-title":"Happily (n)ever after: Aging in the context of oxidative stress, proteostasis loss and cellular senescence","volume":"11","author":"Hoehn","year":"2017","journal-title":"Redox Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.arr.2016.04.009","article-title":"The crucial impact of lysosomes in aging and longevity","volume":"32","author":"Hughes","year":"2016","journal-title":"Ageing Res. Rev."},{"key":"ref_36","first-page":"1005","article-title":"Age-related decline in lysosomal enzyme release from polymorphonuclear leukocytes after N-formyl-methionyl-leucyl-phenylalanine stimulation","volume":"11","author":"Suzuki","year":"1983","journal-title":"Exp. Hematol."},{"key":"ref_37","unstructured":"Lambert, J.R.A. (2018). Less Is More: The Efficacy of Gene Therapy to Treat. Fabry Disease, UCL Library Open Access University College."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5326","DOI":"10.1073\/pnas.76.10.5326","article-title":"Enzyme therapy in Fabry disease: Differential in vivo plasma clearance and metabolic effectiveness of plasma and splenic alpha-galactosidase A isozymes","volume":"76","author":"Desnick","year":"1979","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.ijcme.2016.06.002","article-title":"Myocardial fibrosis pathology in Anderson-Fabry disease: Evaluation of autopsy cases in the long- and short-term enzyme replacement therapy, and non-therapy case","volume":"12","author":"Nagano","year":"2016","journal-title":"IJC Metab. Endocr."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1046\/j.1471-4159.2001.00669.x","article-title":"Local and global cerebral blood flow and glucose utilization in the alpha-galactosidase A knockout mouse model of Fabry disease","volume":"79","author":"Itoh","year":"2001","journal-title":"J. Neurochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"750","DOI":"10.1016\/j.neuint.2013.10.002","article-title":"Levels of 5-methyltetrahydrofolate and ascorbic acid in cerebrospinal fluid are correlated: Implications for the accelerated degradation of folate by reactive oxygen species","volume":"63","author":"Aylett","year":"2013","journal-title":"Neurochem. Int."},{"key":"ref_42","unstructured":"Al Sharani, M. (2018). Mitochondiral function, Oxidative Stress and Parkinson\u2019s Disease, UCL Library Open Access University College."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"7717","DOI":"10.1073\/pnas.87.19.7717","article-title":"Exocytic transport vesicles generated in vitro from the trans-Golgi network carry secretory and plasma membrane proteins","volume":"87","author":"Salamero","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.ymgme.2014.11.005","article-title":"Absence of alpha-galactosidase cross-correction in Fabry heterozygote cultured skin fibroblasts","volume":"114","author":"Fuller","year":"2015","journal-title":"Mol. Genet. Metab."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1042\/BJ20120146","article-title":"Lentiviral vectors: Basic to translational","volume":"443","author":"Sakuma","year":"2012","journal-title":"Biochem. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/0003-2697(85)90442-7","article-title":"Measurement of protein using bicinchoninic acid","volume":"150","author":"Smith","year":"1985","journal-title":"Anal. Biochem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1136\/jmg.18.5.359","article-title":"Antenatal diagnosis of Niemann-Pick disease in a twin pregnancy","volume":"18","author":"Donnai","year":"1981","journal-title":"J. Med. Genet."},{"key":"ref_48","first-page":"491","article-title":"The neurological complications of Anderson-Fabry disease (alpha-galactosidase A deficiency)-investigation of symptomatic and presymptomatic patients","volume":"75","author":"Morgan","year":"1990","journal-title":"Q. J. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/0009-8981(81)90384-3","article-title":"Differential assay for lysosomal alpha-galactosidases in human tissues and its application to Fabry\u2019s disease","volume":"112","author":"Mayes","year":"1981","journal-title":"Clin. Chim. Acta Int. J. Clin. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1136\/jmg.24.5.283","article-title":"Neuraminidase deficiency: Case report and review of the phenotype","volume":"24","author":"Young","year":"1987","journal-title":"J. Med. Genet."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"724","DOI":"10.1007\/BF00712015","article-title":"Mild phenotypic expression of alpha-N-acetylgalactosaminidase deficiency in two adult siblings","volume":"17","author":"Chabas","year":"1994","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1136\/adc.48.10.775","article-title":"Late onset GM2-gangliosidosis. Clinical, pathological, and biochemical studies on 8 patients","volume":"48","author":"Brett","year":"1973","journal-title":"Arch. Dis. Child."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1056\/NEJM197007022830104","article-title":"Tay-sachs disease. Detection of heterozygotes and homozygotes by serum hexosaminidase assay","volume":"283","author":"Okada","year":"1970","journal-title":"N. Engl. J. Med."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"570","DOI":"10.1126\/science.162.3853.570","article-title":"Hurler and Hunter syndromes: Mutual correction of the defect in cultured fibroblasts","volume":"162","author":"Fratantoni","year":"1968","journal-title":"Science"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1111\/j.1471-4159.1990.tb02325.x","article-title":"Mitochondrial complex I deficiency in Parkinson\u2019s disease","volume":"54","author":"Schapira","year":"1990","journal-title":"J. Neurochem."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1042\/bj1140597","article-title":"The kinetic properties of citrate synthase from rat liver mitochondria","volume":"114","author":"Shepherd","year":"1969","journal-title":"Biochem. J."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1111\/jnc.12646","article-title":"The ketogenic diet component increases mitochondrial citrate synthase and complex I activity in neuronal cells","volume":"129","author":"Hughes","year":"2015","journal-title":"J. Neurochem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3349","DOI":"10.1113\/jphysiol.2012.230185","article-title":"Biomarkers of mitochondiral content in skeletal muscle of healthy young subjects","volume":"590","author":"Larsen","year":"2012","journal-title":"J. Physiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/18\/4349\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T13:16:59Z","timestamp":1760188619000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/20\/18\/4349"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,9,5]]},"references-count":58,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2019,9]]}},"alternative-id":["ijms20184349"],"URL":"https:\/\/doi.org\/10.3390\/ijms20184349","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,9,5]]}}}